Dissociated neurons from rat hippocampus are often used to study the kinetics of neural transmission due to their ability to form connections and plasticity. We have developed a protocol using a variety of established methods for the growth and maintenance of hippocampal neurons from 1-5 day old rats. These neurons were imaged using widefield and confocal microscopy. The following features were investigated: a) internal calcium after loading with the AM dye fluo-3. Responses to potassium and glutamate stimulation were observed; b) synaptic vesicle staining using styryl dye FM1-43. Again, responses to potassium and glutamate stimulation were observed; c) synaptic vesicle staining of fixed cells using synapsin I (a protein found at synaptic terminals)- antibody; d) labeling of NADH receptors after loading with conatotoxin; e) testing general microinjection capabilities with FM1-43 and fluorescein-labeled dextran.